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000893282 1001_ $$0P:(DE-HGF)0$$aReinartz, Ines$$b0
000893282 245__ $$aStructural Analysis of a Genetically Encoded FRET Biosensor by SAXS and MD Simulations
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000893282 520__ $$aInspired by the modular architecture of natural signaling proteins, ligand binding proteins are equipped with two fluorescent proteins (FPs) in order to obtain Förster resonance energy transfer (FRET)-based biosensors. Here, we investigated a glucose sensor where the donor and acceptor FPs were attached to a glucose binding protein using a variety of different linker sequences. For three resulting sensor constructs the corresponding glucose induced conformational changes were measured by small angle X-ray scattering (SAXS) and compared to recently published single molecule FRET results (Höfig et al., ACS Sensors, 2018). For one construct which exhibits a high change in energy transfer and a large change of the radius of gyration upon ligand binding, we performed coarse-grained molecular dynamics simulations for the ligand-free and the ligand-bound state. Our analysis indicates that a carefully designed attachment of the donor FP is crucial for the proper transfer of the glucose induced conformational change of the glucose binding protein into a well pronounced FRET signal change as measured in this sensor construct. Since the other FP (acceptor) does not experience such a glucose induced alteration, it becomes apparent that only one of the FPs needs to have a well-adjusted attachment to the glucose binding protein.
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000893282 7001_ $$0P:(DE-Juel1)171618$$aSarter, Mona$$b1
000893282 7001_ $$0P:(DE-Juel1)165764$$aOtten, Julia$$b2
000893282 7001_ $$0P:(DE-Juel1)185813$$aHöfig, Henning$$b3$$ufzj
000893282 7001_ $$0P:(DE-Juel1)131522$$aPohl, Martina$$b4
000893282 7001_ $$0P:(DE-Juel1)173652$$aSchug, Alexander$$b5
000893282 7001_ $$0P:(DE-Juel1)140278$$aStadler, Andreas M.$$b6
000893282 7001_ $$0P:(DE-Juel1)131961$$aFitter, Jörg$$b7$$eCorresponding author
000893282 773__ $$0PERI:(DE-600)2052857-7$$a10.3390/s21124144$$gVol. 21, no. 12, p. 4144 -$$n12$$p4144 -$$tSensors$$v21$$x1424-8220$$y2021
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